title: Time-irreversible subconductance gating associated with Ba2+ block of large conductance Ca2+-activated K+ channels.
abstract: Ba2+ block of large conductance Ca2+-activated K+ channels was studied in patches of membrane excised from cultures of rat skeletal muscle using the patch clamp technique. Under conditions in which a blocking Ba2+ ion would dissociate to the external solution (150 mM N-methyl-D-glucamine+o, 500 mM K+i, 10 microM Ba2+i, +30 mV, and 100 microM Ca2+i to fully activate the channel), Ba2+ blocks with a mean duration of approximately 2 s occurred, on average, once every approximately 100 ms of channel open time. Of these Ba2+ blocks, 78% terminated with a single step in the current to the fully open level and 22% terminated with a transition to a subconductance level at approximately 0.26 of the fully open level (preopening) before stepping to the fully open level. Only one apparent preclosing was observed in approximately 10,000 Ba2+ blocks. Thus, the preopenings represent Ba2+-induced time-irreversible subconductance gating. The fraction of Ba2+ blocks terminating with a preopening and the duration of preopenings (exponentially distributed, mean = 0.75 ms) appeared independent of changes in [Ba2+]i or membrane potential. The fractional conductance of the preopenings increased from 0.24 at +10 mV to 0.39 at +90 mV. In contrast, the average subconductance level during normal gating in the absence of Ba2+ was independent of membrane potential, suggesting different mechanisms for preopenings and normal subconductance levels. Preopenings were also observed with 10 mM Ba2+o and no added Ba2+i. Adding K+, Rb+, or Na+ to the external solution decreased the fraction of Ba2+ blocks with preopenings, with K+ and Rb+ being more effective than Na+. These results are consistent with models in which the blocking Ba2+ ion either induces a preopening gate, and then dissociates to the external solution, or moves to a site located on the external side of the Ba2+ blocking site and acts directly as the preopening gate.
authors.affiliation: Department of Physiology and Biophysics, University of Miami School of Medicine, Miami, Florida 33101-6430, USA.
authors.forename: R A
authors.initials: RA
authors.lastname: Bello
authors.forename: K L
authors.initials: KL
authors.lastname: Magleby
mesh_terms: D000818:Animals; D001464:Barium / Q000494:pharmacology*; D002118:Calcium / Q000502:physiology*; D002478:Cells, Cultured; D004594:Electrophysiology; D005260:Female; D015640:Ion Channel Gating / Q000187:drug effects*; D008564:Membrane Potentials / Q000502:physiology; D018482:Muscle, Skeletal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D018408:Patch-Clamp Techniques; D026902:Potassium Channel Blockers; D015221:Potassium Channels / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors
chemical_list: D026902:Potassium Channel Blockers; D015221:Potassium Channels; D001464:Barium; D002118:Calcium